为了探讨玉米赤霉烯酮(ZEA)与脱氧雪腐镰刀菌烯醇(DON)联合作用对免疫系统毒性影响及其毒理机制,试验以T淋巴细胞株(CTLL-2)为材料,在利用CCK-8试验确定ZEA、DON对CTLL-2细胞联合染毒浓度的基础上,研究了ZEA与DON联合染毒对CTLL-2细胞...为了探讨玉米赤霉烯酮(ZEA)与脱氧雪腐镰刀菌烯醇(DON)联合作用对免疫系统毒性影响及其毒理机制,试验以T淋巴细胞株(CTLL-2)为材料,在利用CCK-8试验确定ZEA、DON对CTLL-2细胞联合染毒浓度的基础上,研究了ZEA与DON联合染毒对CTLL-2细胞凋亡及凋亡相关蛋白Bax、Bcl-2和Caspase-3等表达的影响,试验设对照组、ZEA染毒组(5μg/m L ZEA)、DON染毒组(0.5μg/m L DON)、ZEA+DON联合染毒组(5μg/m L ZEA+0.5μg/m L DON),染毒时间为48 h,用流式细胞术检测细胞的凋亡率,Western-blot检测Bax、Bcl-2和Caspase级联反应相关蛋白的表达情况。结果表明:与对照组相比,各染毒组CTLL-2细胞的凋亡率均极显著升高(P<0.01);各染毒组细胞凋亡相关蛋白Bax/Bcl-2比值和Cleaved Cspase-3、Cleaved Caspase-8、Cleaved Caspase-9等蛋白的表达量均较对照组显著升高(P<0.05或P<0.01);联合染毒组呈现协同效应。说明ZEA与DON联合染毒在诱导免疫细胞凋亡方面发挥协同效应,它们共同存在可以发挥更强的免疫抑制效应;ZEA与DON联合染毒诱导CTLL-2细胞发生凋亡的机制与Bax/Bcl-2上调的线粒体通路密切相关。展开更多
目的分析腹腔镜胆囊切除术(LC)对胆结石患者血清胆囊收缩素A型受体(CCK-A)、固醇调节元件结合蛋白2(SREBP-2)及胆红素(TCB)水平的影响。方法选取2017年6月至2019年4月我院收治的胆结石患者140例,依据非随机临床同期对照研究及患者自愿...目的分析腹腔镜胆囊切除术(LC)对胆结石患者血清胆囊收缩素A型受体(CCK-A)、固醇调节元件结合蛋白2(SREBP-2)及胆红素(TCB)水平的影响。方法选取2017年6月至2019年4月我院收治的胆结石患者140例,依据非随机临床同期对照研究及患者自愿原则分为观察组合和对照组各70例。对照组予以小切口胆囊切除术(MC),观察组予以LC术,比较两组手术一般情况、手术前后血清CCK-A、SREBP-2、TCB、疼痛(简化McGill疼痛问卷)、并发症发生率。结果观察组手术时间、术后住院时间、肠鸣音复常时间、下地活动时间均短于对照组,术中出血量较对照组低(P<0.05);观察组术后CCK-A水平高于对照组,SREBP-2、TCB低于对照组(P<0.05);观察组术后1、3、7 d McGill评分低于对照组(P<0.05),术后1个月内并发症发生率低于对照组(P<0.05)。结论与MC术相比,LC术治疗胆结石疗效更好,可有效促进患者术后恢复,提高CCK-A水平,下调SREBP-2、TCB及炎症因子水平,降低疼痛和并发症风险,值得临床推广实践。展开更多
Objective： The aim of the study was to investigate the apoptosis induced by piperlongumine on human breast adenoma MDA-MB-231 cells and the mechanism involved. Methods： Human breast adenoma MDA-MB-231 cells line was...Objective： The aim of the study was to investigate the apoptosis induced by piperlongumine on human breast adenoma MDA-MB-231 cells and the mechanism involved. Methods： Human breast adenoma MDA-MB-231 cells line was cultured in vitro. The inhibitory effect of piperlongumine on the proliferation of human breast adenoma MDA-MB-231 cells was measured by CCK-8 assay. Distribution of cell cycle was analyzed by flow cytometry. The apoptosis rates of MDA-MB- 231 cells were measured using Annexin V/PI staining. The flow cytometry with the probe of DCFH-DA was used to detect the intracellular reactive oxygen species levels. Western blot was used to explore the protein expression of Bcl-2 and Bax. Results： The CCK-8 assay showed that piperlongumine had an inhibiting effect on the proliferation of MDA-MB-231 cells in a concentrationand time-dependent manner. MDA-MB-231 cells were markedly arrested at G0/G1 phase after treatment of piperlongumine. Piperlongumine induced apoptosis of MDA-MB-231 cells obviously. The level of intracellular reactive oxygen species was increased in a dose-dependent manner. The antioxidant N-acetyI-L-cystein inhibited the apoptosis of cells and the level of intracellular reactive oxygen species was also decreased. By Western blot analysis, we found the expression of Bax was up-regulated whereas that of Bcl-2 was down-regulated in a concentration-dependent manner. Conclusion： PiperIongumine possesses a significant function for inhibiting proliferation, arresting cells at G0/G1 phase and inducing apoptosis of MDA-MB-231 cells, which seems to be associated with the increased generation of intracellular reactive oxygen species as well as the down-regulation of Bcl-2 and up-regulation of Bax.展开更多
AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confi...AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confirmed by real-time reverse transcription- polymerase chain reaction （PCR）. Western blotting and semi-quantitative immunohistochemistry were used to examine protein levels in human pancreatic cancer cell lines and pancreatic tissues, respectively, The role of αv integrin on gastrin-induced cell adhesion was examined using blocking anti-αv integrin monoclonal antibodies. Adherent cells were quantified by staining with crystal violet.RESULTS： Using a ＂cancer genes＂ array we identi- fied c^v integrin as a new gastrin target gene in human pancreatic cancer cells. A quantitative real-time PCR approach was used to confirm αv integrin gene expression. We also demonstrate that Src family kinases and the PI 3-kinase, two signalling pathways specifically activated by the CCK-2 receptor （CCK2R）, are involved in gastrin-mediated αv integrin expression. In contrast, inhibition of the ERK pathway was without any effect on αv integrin expression induced by gastrin. Our results also show that gastrin modulates cell adhesion via αv integrins. Indeed, in vitro adhesion assays performed on fibronectin show that gastrin significantly increases adhesion of pancreatic cancer cells. The use of blocking anti-αv integrin monoclonal antibodies completely reversed the increase in cell-substrate adhesion induced by gastrin. In addition, we showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, leads to the overexpression of αv integrin. This process may contribute to pancreatic turnout development observed in these transgenic animals.CONCLUSION： αv integrin is a new gastrin target in pancreatic cancer models and contributes to gastrin effects on cell adhesion.展开更多
Objective Probe into the influence and the mechanisms of CCK and gastrin on the apoptosis of bile duct carcinoma cells. Methods By taking beauvericin as the revulsant to the apoptosis of bile duct carcinoma cells, the...Objective Probe into the influence and the mechanisms of CCK and gastrin on the apoptosis of bile duct carcinoma cells. Methods By taking beauvericin as the revulsant to the apoptosis of bile duct carcinoma cells, the influence of CCK and gastrin on the apoptosis of bile duct carcinoma cells was investigated by using the techniques such as TUNEL fluorescent staining, stream mode cell detecting instrument and reverse bcl-2 oligonucleotide. Techniques of immunohistochemistry, in situ hybridization, flow cytometry (FCM) , RT-PCR were used to study the roles of apoptosis-related genes bcl-2 and baxResults After beauvericin 40 uM worked for 12 h, the survival rate of QBC939 bile duct carcinoma cells was decreased by 35% ?40% . About 80% of the bile duct carcinoma cells showed various degrees of apoptosis. CCK and gastrin could upregulate the threshold value of the apoptosis of bile duct carcinoma cells, which could be inhibited by L60, L18 and reverse bcl-2 oligonucleotide. In terms of both transcription and translating levels, CCK and gastrin could obviously promote the genetic expression of bcl-2, but had no influence on the genetic expression of bax. Addition of CCK-A receptor or CCK-B/gastr in receptor antagonist could remarkably inhibit the expression of bcl-2 boosted by gastrin-17 and CCK-8S.Conclusion CCK and gastrin inhibited the apoptosis of bile duct carcinoma cells through upregulating the genetic expression of bcl-2. Theoretically, this research has expanded our understanding to the mechanism of CCK and gastrin in controlling the growth of tumors, enriched our view to the mechanism of apoptosis of alimentary tract tumors, and has provided a new thinking for the assistant treatment to bile duct carcinoma cells as well.展开更多
文摘为了探讨玉米赤霉烯酮(ZEA)与脱氧雪腐镰刀菌烯醇(DON)联合作用对免疫系统毒性影响及其毒理机制,试验以T淋巴细胞株(CTLL-2)为材料,在利用CCK-8试验确定ZEA、DON对CTLL-2细胞联合染毒浓度的基础上,研究了ZEA与DON联合染毒对CTLL-2细胞凋亡及凋亡相关蛋白Bax、Bcl-2和Caspase-3等表达的影响,试验设对照组、ZEA染毒组(5μg/m L ZEA)、DON染毒组(0.5μg/m L DON)、ZEA+DON联合染毒组(5μg/m L ZEA+0.5μg/m L DON),染毒时间为48 h,用流式细胞术检测细胞的凋亡率,Western-blot检测Bax、Bcl-2和Caspase级联反应相关蛋白的表达情况。结果表明:与对照组相比,各染毒组CTLL-2细胞的凋亡率均极显著升高(P<0.01);各染毒组细胞凋亡相关蛋白Bax/Bcl-2比值和Cleaved Cspase-3、Cleaved Caspase-8、Cleaved Caspase-9等蛋白的表达量均较对照组显著升高(P<0.05或P<0.01);联合染毒组呈现协同效应。说明ZEA与DON联合染毒在诱导免疫细胞凋亡方面发挥协同效应,它们共同存在可以发挥更强的免疫抑制效应;ZEA与DON联合染毒诱导CTLL-2细胞发生凋亡的机制与Bax/Bcl-2上调的线粒体通路密切相关。
文摘目的分析腹腔镜胆囊切除术(LC)对胆结石患者血清胆囊收缩素A型受体(CCK-A)、固醇调节元件结合蛋白2(SREBP-2)及胆红素(TCB)水平的影响。方法选取2017年6月至2019年4月我院收治的胆结石患者140例,依据非随机临床同期对照研究及患者自愿原则分为观察组合和对照组各70例。对照组予以小切口胆囊切除术(MC),观察组予以LC术,比较两组手术一般情况、手术前后血清CCK-A、SREBP-2、TCB、疼痛(简化McGill疼痛问卷)、并发症发生率。结果观察组手术时间、术后住院时间、肠鸣音复常时间、下地活动时间均短于对照组,术中出血量较对照组低(P<0.05);观察组术后CCK-A水平高于对照组,SREBP-2、TCB低于对照组(P<0.05);观察组术后1、3、7 d McGill评分低于对照组(P<0.05),术后1个月内并发症发生率低于对照组(P<0.05)。结论与MC术相比,LC术治疗胆结石疗效更好,可有效促进患者术后恢复,提高CCK-A水平,下调SREBP-2、TCB及炎症因子水平,降低疼痛和并发症风险,值得临床推广实践。
基金Supported by grants from the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. JX10231801)the Innovative Medical Team and Leading Talent of Jiangsu Province (No. LJ201123)
文摘Objective： The aim of the study was to investigate the apoptosis induced by piperlongumine on human breast adenoma MDA-MB-231 cells and the mechanism involved. Methods： Human breast adenoma MDA-MB-231 cells line was cultured in vitro. The inhibitory effect of piperlongumine on the proliferation of human breast adenoma MDA-MB-231 cells was measured by CCK-8 assay. Distribution of cell cycle was analyzed by flow cytometry. The apoptosis rates of MDA-MB- 231 cells were measured using Annexin V/PI staining. The flow cytometry with the probe of DCFH-DA was used to detect the intracellular reactive oxygen species levels. Western blot was used to explore the protein expression of Bcl-2 and Bax. Results： The CCK-8 assay showed that piperlongumine had an inhibiting effect on the proliferation of MDA-MB-231 cells in a concentrationand time-dependent manner. MDA-MB-231 cells were markedly arrested at G0/G1 phase after treatment of piperlongumine. Piperlongumine induced apoptosis of MDA-MB-231 cells obviously. The level of intracellular reactive oxygen species was increased in a dose-dependent manner. The antioxidant N-acetyI-L-cystein inhibited the apoptosis of cells and the level of intracellular reactive oxygen species was also decreased. By Western blot analysis, we found the expression of Bax was up-regulated whereas that of Bcl-2 was down-regulated in a concentration-dependent manner. Conclusion： PiperIongumine possesses a significant function for inhibiting proliferation, arresting cells at G0/G1 phase and inducing apoptosis of MDA-MB-231 cells, which seems to be associated with the increased generation of intracellular reactive oxygen species as well as the down-regulation of Bcl-2 and up-regulation of Bax.
文摘AIM： To analyse αv integrin expression induced by gas- trin in pancreatic cancer models.METHODS： αv integrin mRNA expression in human pan- creatic cancer cells was analysed using a ＂cancer genes＂ array and confirmed by real-time reverse transcription- polymerase chain reaction （PCR）. Western blotting and semi-quantitative immunohistochemistry were used to examine protein levels in human pancreatic cancer cell lines and pancreatic tissues, respectively, The role of αv integrin on gastrin-induced cell adhesion was examined using blocking anti-αv integrin monoclonal antibodies. Adherent cells were quantified by staining with crystal violet.RESULTS： Using a ＂cancer genes＂ array we identi- fied c^v integrin as a new gastrin target gene in human pancreatic cancer cells. A quantitative real-time PCR approach was used to confirm αv integrin gene expression. We also demonstrate that Src family kinases and the PI 3-kinase, two signalling pathways specifically activated by the CCK-2 receptor （CCK2R）, are involved in gastrin-mediated αv integrin expression. In contrast, inhibition of the ERK pathway was without any effect on αv integrin expression induced by gastrin. Our results also show that gastrin modulates cell adhesion via αv integrins. Indeed, in vitro adhesion assays performed on fibronectin show that gastrin significantly increases adhesion of pancreatic cancer cells. The use of blocking anti-αv integrin monoclonal antibodies completely reversed the increase in cell-substrate adhesion induced by gastrin. In addition, we showed in vivo that the targeted CCK2R expression in the pancreas of Elas-CCK2 mice, leads to the overexpression of αv integrin. This process may contribute to pancreatic turnout development observed in these transgenic animals.CONCLUSION： αv integrin is a new gastrin target in pancreatic cancer models and contributes to gastrin effects on cell adhesion.
基金Supported by National Natural Science Foundation of China.Project number:39670711
文摘Objective Probe into the influence and the mechanisms of CCK and gastrin on the apoptosis of bile duct carcinoma cells. Methods By taking beauvericin as the revulsant to the apoptosis of bile duct carcinoma cells, the influence of CCK and gastrin on the apoptosis of bile duct carcinoma cells was investigated by using the techniques such as TUNEL fluorescent staining, stream mode cell detecting instrument and reverse bcl-2 oligonucleotide. Techniques of immunohistochemistry, in situ hybridization, flow cytometry (FCM) , RT-PCR were used to study the roles of apoptosis-related genes bcl-2 and baxResults After beauvericin 40 uM worked for 12 h, the survival rate of QBC939 bile duct carcinoma cells was decreased by 35% ?40% . About 80% of the bile duct carcinoma cells showed various degrees of apoptosis. CCK and gastrin could upregulate the threshold value of the apoptosis of bile duct carcinoma cells, which could be inhibited by L60, L18 and reverse bcl-2 oligonucleotide. In terms of both transcription and translating levels, CCK and gastrin could obviously promote the genetic expression of bcl-2, but had no influence on the genetic expression of bax. Addition of CCK-A receptor or CCK-B/gastr in receptor antagonist could remarkably inhibit the expression of bcl-2 boosted by gastrin-17 and CCK-8S.Conclusion CCK and gastrin inhibited the apoptosis of bile duct carcinoma cells through upregulating the genetic expression of bcl-2. Theoretically, this research has expanded our understanding to the mechanism of CCK and gastrin in controlling the growth of tumors, enriched our view to the mechanism of apoptosis of alimentary tract tumors, and has provided a new thinking for the assistant treatment to bile duct carcinoma cells as well.
